Cholera vaccine

Abstract

A particulate material for inducing immunity to cholera toxin comprises support particles provided with a first coating of a ganglioside having affinity for cholera toxin and a second coating of cholera toxin attached to said ganglioside coating. This particulate material can be used as an orally or parenterally administered medicine for humans to induce an immunity response to cholera toxin.

We claim: 
     
       1. A process for the preparation of a particulate material capable of inducing immunity and inducing the synthesis of antibodies to cholera toxin when administered to a living organism that possesses an immunity defense system which comprises support particles provided with a first coating of a ganglioside having an affinity for cholera toxin and a second coating of cholera toxin attached to said ganglioside coating, said support particles being selected from red blood corpuscles, inactivated bacteria particles, an organic polymer capable of giving a stable suspension in water, carbon particles or porous mineral particles coated with a polysaccharide polymer or with an oxidative scission product thereof, crosslinked or not, said process comprising in a first stage affixing said ganglioside onto said support particles to provide said first coating on said support particles and in a second stage affixing said cholera toxin onto said ganglioside coating. 
     
     
       2. The process of claim 1 wherein the second stage is effected by contacting the ganglioside coated particles with cholera toxin for a time ranging from 1 to 60 minutes at a temperature ranging from 0° to 60° C. 
     
     
       3. The process of claim 1 wherein the support particles are red blood corpuscles treated with formaldehyde or glutaraldehyde. 
     
     
       4. The process of claim 3 wherein the treatment of said red blood corpuscles comprises reacting said red blood corpuscles in suspension in a physiological salt solution with a physiological salt solution of formaldehyde or glutaraldehyde for a period of 15 minutes to 15 hours at a temperature ranging from 0° to 60° C., the formaldehyde or glutaraldehyde being present in an amount ranging from 0.01 to 20 grams per 100 ml of red blood corpuscle suspension. 
     
     
       5. The process of claim 4 wherein the thus treated red blood corpuscles are contacted with a suspension of ganglioside G M1  at a pH equal to or greater than 8 and at an incubation temperature ranging from 0° to 60° C. for an incubation time ranging from about 1 hour to 15 hours. 
     
     
       6. The process of claim 5 wherein the pH is about 13, the temperature is about 45° C. and the time is about 15 hours. 
     
     
       7. The process of claim 4 wherein the thus treated red blood corpuscles are contacted with a suspension of lysoganglioside G M1  at a pH above 5 and at an incubation temperature ranging from 0° to 60° C. for an incubation time ranging from 30 minutes to 30 hours. 
     
     
       8. The process of claim 7 wherein the pH is about 7. 
     
     
       9. The process of claim 1 wherein the support particle is an inactivated bacteria particle. 
     
     
       10. The process of claim 9 wherein the bacteria particle is cholera vibrios. 
     
     
       11. The process of claim 1 wherein the support particle is an organic polymer capable of giving a stable suspension in water. 
     
     
       12. The process of claim 11 wherein said organic polymer is styrene, butadiene or divinylbenzene. 
     
     
       13. The process of claim 1 wherein the support particle is a carbon particle. 
     
     
       14. The process of claim 1 wherein the support particle is a porous mineral particle coated with an aminopolysaccharide or an oxidative scission product thereof, crosslinked or not. 
     
     
       15. A process for the preparation of a particulate material capable of inducing immunity and inducing the synthesis of antibodies to cholera toxin when administered to a living organism that possesses an immunity defense system which comprises support particles provided with a first coating of a ganglioside having an affinity for cholera toxin and a second coating of cholera toxin attached to said ganglioside coating, said support particles being selected from red blood corpuscles, inactivated bacteria particles, an organic polymer capable of giving a stable suspension in water, carbon particles or porous mineral particles coated with a polysaccharide polymer or with an oxidative scission product thereof, crosslinked or not comprising in a first stage affixing said ganglioside onto said support particles to provide said first coating on said support particles and in a second stage attaching said cholera toxin onto said ganglioside coating, the amount of attached cholera toxin being such that the attachment sites of said ganglioside are not saturated with said cholera toxin. 
     
     
       16. An parenterally administrable medicine for inducing an immunity response to cholera toxin comprising a dosage of a particulate material capable of inducing immunity and inducing the synthesis of antibodies to cholera toxin consisting essentially of support particles selected from inactivated bacteria particles and soluble polysaccharides provided with a first coating of a ganglioside having an affinity for cholera toxin and a second coating of cholera toxin attached to said ganglioside coating, said dosage being effective to induce said immunity response. 
     
     
       17. A medicine for inducing an immunity response to cholera toxin comprising a dosage of a particulate material capable of inducing immunity and inducing the synthesis of antibodies to cholera when administered to a living organism that possesses an immunity defense system which consists essentially of support particles provided with a first coating of a ganglioside having an affinity for cholera toxin and a second coating of cholera toxin attached to said ganglioside coating, said dosage being effective to induce said immunity response. 
     
     
       18. The medicine of claim 17 wherein said support particles are selected from red blood corpuscles, polysaccharide particles, inactivated bacteria particles, an organic polymer capable of giving a stable suspension in water, carbon particles or porous mineral particles coated with a polysaccharide polymer or with an oxidative scission product thereof, crosslinked or not. 
     
     
       19. The medicine of claim 17 for oral administration wherein the amount of cholera toxin attached to said first coating is such that the attachment sites of said ganglioside are not saturated with said cholera toxin. 
     
     
       20. The medicine of claim 17 wherein the support particles are red blood corpuscles. 
     
     
       21. The medicine of claim 17 wherein the support particles are inactivated bacteria particles. 
     
     
       22. The medicine of claim 21 wherein the inactivated bacteria particles are cholera vibrios. 
     
     
       23. The medicine of claim 17 wherein the support particles are an organic polymer selected from styrene, butadiene and divinylbenzene. 
     
     
       24. The medicine of claim 17 wherein the support particles are carbon particles. 
     
     
       25. The medicine of claim 17 wherein the support particles are polysaccharide particles. 
     
     
       26. The medicine of claim 25 wherein the polysaccharide is an aminopolysaccharide. 
     
     
       27. The medicine of claim 17 wherein the support particles are porous mineral particles coated with an aminopolysaccharide polymer or an oxidative scission product thereof, crosslinked or not. 
     
     
       28. A process for inducing the formation of cholera antitoxin antibodies comprising injecting into an animal a particulate material comprising support particles provided with a first coating of a ganglioside having an affinity for cholera toxin and a second coating of cholera toxin attached to said ganglioside coating, said support particles being selected from red blood corpuscles, inactivated bacteria particles, an organic polymer capable of giving a stable suspension in water, carbon particles or porous mineral particles coated with a polysaccharide polymer or with an oxidative scission product thereof, crosslinked or not so as to induce the synthesis of said cholera antitoxin antibodies and subsequently isolating the said resulting antibodies. 
     
     
       29. A process for inducing an immunity response to cholera toxin comprising orally administering to a living organism possessing an immunity defense system the medicine of claim 17 in a dosage amount effective to induce said immunity response. 
     
     
       30. A process for inducing an immunity response to cholera toxin comprising parenterally administering to a vibrio carrying human the medicine of claim 16 in a dosage amount effective to induce said immunity response. 
     
     
       31. A process for inducing an immunity response to cholera toxin comprising parenterally administering to a living organism possessing an immunity defense system the medicine of claim 17 in a dosage amount effective to induce said immunity response, wherein the support particle is an inactivated bacteria particle or a polysaccharide.